The Value of CTA Based on Gold Nanorod Contrast Agent in Coronary Artery Diagnosis and Plaque Property Analysis.

Coronary CT angiography (CTA) with the characteristics of noninvasive and simple operation is widely used in the diagnosis of coronary artery stenosis. The choice of contrast agent exerts an important impact on the imaging quality of CTA. Conventional iodine contrast agents are easily excreted by the kidneys, from which the imaging window is short, and the imaging quality is poor. Metal nanomaterials have unique optical properties and have broad application prospects in imaging. Our aim is to explore the value of gold nanorod contrast agent in the diagnosis of coronary heart disease. A gold nanorod suspension was first prepared, and the prepared gold nanorod was uniform and had good dispersibility. It can be seen from the light absorption curve that there are two obvious peaks on the UV absorption peak of the gold nanorods. The gold nanorods were cultured in different solutions, and it was found that the particle size of the gold nanorods did not change significantly within 72 hours, indicating that the prepared gold nanorods had good stability. When observing the damage degree of mouse kidney tissue, it was shown that the damage degree of gold nanorod contrast agent to mouse kidney tissue was less than that of iodine contrast agent. The above results indicate that the gold nanorod contrast agent has good stability and safety. Therefore, our study demonstrated that the gold nanorod contrast agent has high value in the diagnosis of coronary arteries and the analysis of plaque properties.

Design and characterization of nanocarriers loaded with Levofloxacin for enhanced antimicrobial activity; physicochemical properties, in vitro release and oral acute toxicity

Inorganic and carbon based nanomaterials are widely used against several diseases, such as cancer, autoimmune diseases as well as fungi and bacteria colonization. In this work, Santa Barbara Amorphous mesoporous silica (SBA), Halloysite Nanotubes (HNTs) and Multiwalled Carbon Nanotubes (CNTs) were loaded with fluoroquinolone Levofloxacin (LVF) to be applied as antimicrobial agents. The prepared via adsorption nanocarriers were characterized by Fourier-Transformed Spectroscopy, Scanning Electron Microscopy as well as High Pressure liquid Chromatography. In vitro release studies were carried out using Simulated Body Fluid at 37oC and data analyzed by various kinetic models showing slow dissolution over 12-24 hours. Antimicrobial studies showed improved antibacterial activity against Escherichia coli, Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus, and Staphylococcus epidermidis compared to neat nanomaterials. CNTs were found to be the most promising candidates for LVF delivery and they were chosen to be further studied for their acute oral toxicity and histopathological examination using C57/Black mice. Histological examination depicted that drug loading did not affect mice organs morphology as well as hepatocyte degeneration, central vein degeneration and parenchymal necrosis scores. To conclude, the prepared nanomaterials present significant characteristics and can act as antimicrobial drug carriers; CNTs found to be safe candidates when orally fed to mice.

Catechin loaded PLGA submicron-sized fibers reduce levels of reactive oxygen species induced by MWCNT in vitro.

Reactive oxygen species (ROS) are common products of normal aerobic cellular metabolism, but high levels of ROS lead to oxidative stress and cellular damage. Therefore, effective antioxidant therapies are needed to prevent ROS overproduction. This study reports the development of poly(l-lactide-co-glycolide) (PLGA) bicomponent fibers loaded with selected amounts of the natural polyphenolic antioxidant catechin. Thereby a novel route based on emulsion electrospinning is investigated to obtain tailored and sustained release rates for chatechin. The activity of the released catechin was assessed for its influence on multi-walled carbon nanotube (MWCNT) induced formation of reactive oxygen species (ROS) in the human alveolar epithelial the cell line A549. Homogenous fiber morphologies were obtained at specified ranges of PLGA concentrations within the emulsions including the formation of a core - sheath structure localizing the drug within the fiber core. In vitro measurements of the delivery showed moderate burst release kinetics in a first phase followed by a linear and smooth release at long term. In combination with polymer degradation studies a mostly diffusion controlled release mechanism was revealed exhibiting only marginal degradation of the polymer during the time span of the drug delivery. As a proof of concept, the activity of released catechin in A549 cells stimulated with MWCNTs was determined and revealed a high reduction of ROS production in a dose dependent manner. This effect diminishes over time indicating a depletion of catechin.

Foreign Body Reaction to Biomaterial Nanotubular Surface and the Influence of Silver Loading.

The impacts of biomaterial surface properties (i.e., surface nanotopography) and dopants (i.e., silver (Ag)) on the biomaterial-associated foreign body reaction (FBR) remain unclear. In this study, an in vivo FBR that was induced by a titania nanotube array (NT) on titanium was examined with and without Ag loading. An NT with an 80 nm diameter that was fabricated by anodization, and the NT samples that were loaded with two Ag concentrations (NT-AgH and NT-AgL) and formed by electrodeposition exhibited high hydrophilicities. A relatively rapid initial Ag+ release with a subsequent gradual reduction was observed for NT-AgH and NT-AgL; the Ag+ release was higher for NT-AgH than for NT-AgL. We found that the NT decreased the biomaterial-associated FBR, evidenced by a decreased fibrous capsule thickness and a number of recruited macrophages. The effect of Ag loading on the FBR was considered acceptable because the FBR induced by the NT-Ag samples was still less severe than that of the PT control. Additionally, the temporary increases in the blood, brain, liver and kidney Ag+ concentrations did not produce general side effects and were considered to be in the safe range. This study demonstrates the ability of nanotopography to alleviate the biomaterial-associated FBR and provides further evidence for future clinical applications of biomaterials with nanostructures and Ag loading.

Silicon-Doped Titanium Dioxide Nanotubes Promoted Bone Formation on Titanium Implants.

While titanium (Ti) implants have been extensively used in orthopaedic and dental applications, the intrinsic bioinertness of untreated Ti surface usually results in insufficient osseointegration irrespective of the excellent biocompatibility and mechanical properties of it. In this study, we prepared surface modified Ti substrates in which silicon (Si) was doped into the titanium dioxide (TiO2) nanotubes on Ti surface using plasma immersion ion implantation (PIII) technology. Compared to TiO2 nanotubes and Ti alone, Si-doped TiO2 nanotubes significantly enhanced the expression of genes related to osteogenic differentiation, including Col-I, ALP, Runx2, OCN, and OPN, in mouse pre-osteoblastic MC3T3-E1 cells and deposition of mineral matrix. In vivo, the pull-out mechanical tests after two weeks of implantation in rat femur showed that Si-doped TiO2 nanotubes improved implant fixation strength by 18% and 54% compared to TiO2-NT and Ti implants, respectively. Together, findings from this study indicate that Si-doped TiO2 nanotubes promoted the osteogenic differentiation of osteoblastic cells and improved bone-Ti integration. Therefore, they may have considerable potential for the bioactive surface modification of Ti implants.

Cellulose Nanofibril Hydrogel Tubes as Sacrificial Templates for Freestanding Tubular Cell Constructs.

The merging of defined nanoscale building blocks with advanced additive manufacturing techniques is of eminent importance for the preparation of multiscale and highly functional materials with de novo designed hierarchical architectures. Here, we demonstrate that hydrogels of cellulose nanofibrils (CNF) can be processed into complex shapes, and used as a sacrificial template to prepare freestanding cell constructs. We showcase our approach for the fabrication of hollow fibers using a controlled extrusion through a circular die into a coagulation bath. The dimensions of the hollow fibers are tunable, and the final tubes combine the nanofibrillar porosity of the CNF hydrogel with a submillimeter wall thickness and centimeter-scale length provided by the additive manufacturing technique. We demonstrate that covalent and supramolecular cross-linking of the CNFs can be used to tailor the mechanical properties of the hydrogel tubes within 1 order of magnitude and in an attractive range for the mechanosensation of cells. The resulting tubes are highly biocompatible and allow for the growth of mouse fibroblasts into confluent cell layers in their inner lumen. A detailed screening of several cellulases enables degradation of the scaffolding, temporary CNF hydrogel tube in a quick and highly cell-friendly way, and allows the isolation of coherent cell tubes. We foresee that the growing capabilities of hydrogel printing techniques in combination with the attractive features of CNFs-sustainable, globally abundant, biocompatible and enzymatically degradable-will allow making plant-based biomaterials with hierarchical structures and on-demand degradation useful, for instance, to engineer complex tissue structures to replace animal models, and for implants.

Interactions of 1D- and 2D-layered inorganic nanoparticles with fibroblasts and human mesenchymal stem cells.

AIM: This study investigates the effects of tungsten disulfide nanotubes (WSNTs) and molybdenum disulfide nanoplatelets (MSNPs) on fibroblasts (NIH-3T3) and mesenchymal stem cells (MSCs) to determine safe dosages for potential biomedical applications. MATERIALS & METHODS: Cytotoxicity of MSNPs and WSNTs (5-300 µg/ml) on NIH-3T3 and MSCs was assessed at 6, 12 or 24 h. MSC differentiation to adipocytes and osteoblasts was assessed following treatment for 24 h. RESULTS: Only NIH-3T3 cells treated with MSNPs showed dose or time dependent increase in cytotoxicity. Differentiation markers of MSCs in treated groups were unaffected compared with untreated controls. CONCLUSION: MSNPs and WSNTs at concentrations less than 50 µg/ml are potentially safe for treatment of fibroblasts or MSCs for up to 24 h.

Antibacterial Effects and Biocompatibility of Titania Nanotubes with Octenidine Dihydrochloride/Poly(lactic-co-glycolic acid).

Titanium (Ti) implants with long-term antibacterial ability and good biocompatibility are highly desirable materials that can be used to prevent implant-associated infections. In this study, titania nanotubes (TNTs) were synthesized on Ti surfaces through electrochemical anodization. Octenidine dihydrochloride (OCT)/poly(lactic-co-glycolic acid) (PLGA) was infiltrated into TNTs using a simple solvent-casting technique. OCT/PLGA-TNTs demonstrated sustained drug release and maintained the characteristic hollow structures of TNTs. TNTs (200 nm in diameter) alone exhibited slight antibacterial effect and good osteogenic activity but also evidently impaired adhesion and proliferation of bone marrow mesenchymal stem cells (BMSCs). OCT/PLGA-TNTs (100 nm in diameter) supported BMSC adhesion and proliferation and showed good osteogenesis-inducing ability. OCT/PLGA-TNTs also exhibited good long-term antibacterial ability within the observation period of 7 d. The synthesized drug carrier with relatively long-term antibacterial ability and enhanced excellent biocompatibility demonstrated significant potential in bone implant applications.

Role of signal transducer and activator of transcription 1 in murine allergen-induced airway remodeling and exacerbation by carbon nanotubes.

Asthma is characterized by a T helper type 2 phenotype and by chronic allergen-induced airway inflammation (AAI). Environmental exposure to air pollution ultrafine particles (i.e., nanoparticles) exacerbates AAI, and a concern is possible exacerbation posed by engineered nanoparticles generated by emerging nanotechnologies. Signal transducer and activator of transcription (STAT) 1 is a transcription factor that maintains T helper type 1 cell development. However, the role of STAT1 in regulating AAI or exacerbation by nanoparticles has not been explored. In this study, mice with whole-body knockout of the Stat1 gene (Stat1(-/-)) or wild-type (WT) mice were sensitized to ovalbumin (OVA) allergen and then exposed to multiwalled carbon nanotubes (MWCNTs) by oropharygneal aspiration. In Stat1(-/-) and WT mice, OVA increased eosinophils in bronchoalveolar lavage fluid, whereas MWCNTs increased neutrophils. Interestingly, OVA sensitization prevented MWCNT-induced neutrophilia and caused only eosinophilic inflammation. Stat1(-/-) mice displayed increased IL-13 in bronchoalveolar lavage fluid at 1 day compared with WT mice after treatment with OVA or OVA and MWCNTs. At 21 days, the lungs of OVA-sensitized Stat1(-/-) mice displayed increased eosinophilia, goblet cell hyperplasia, airway fibrosis, and subepithelial apoptosis. MWCNTs further increased OVA-induced goblet cell hyperplasia, airway fibrosis, and apoptosis in Stat1(-/-) mice at 21 days. These changes corresponded to increased levels of profibrogenic mediators (transforming growth factor-ß1, TNF-α, osteopontin) but decreased IL-10 in Stat1(-/-) mice. Finally, fibroblasts isolated from the lungs of Stat1(-/-) mice produced significantly more collagen mRNA and protein in response to transforming growth factor-ß1 compared with WT lung fibroblasts. Our results support a protective role for STAT1 in chronic AAI and exacerbation of remodeling caused by MWCNTs.

Gold nanorods and curcumin-loaded nanomicelles for efficient in vivo photothermal therapy of Barrett's esophagus.

AIM: Provide an enhanced local drug delivery, nanoparticle(s) to minimize systemic effects and achieve enhanced permeability and drug retention into abnormal cells and stroma. MATERIALS & METHODS: Here a simultaneous loading of lipophilic gold nanorods (GNRs) and curcumin into polymeric nanomicelles made of biocompatible PLGA-b-PEG copolymer through a double re-emulsification process has been developed. RESULTS: Initial results in vitro on Barrett's esophagus and esophageal adenocarcinoma cell lines demonstrated a significant reduction in cell viability with curcumin and GNRs exposure (p < 0.05). In vivo Barrett's-associated animal model confirmed these results with successful in vivo demonstrated eradication of all high-grade dysplastic premalignant cancer cells. CONCLUSION: The synthesis of this novel nanosystem containing GNRs and curcumin is safe and effective in treating and eradicating premalignant esophageal adenocarcinoma.

Investigation of the cytotoxic effects of titanate nanotubes on Caco-2 cells.

Titanate nanotubes can be used as drug delivery systems, but limited information is available on their interactions with intestinal cells. In this study, we investigated the cytotoxicity and cellular uptake of titanate nanotubes on Caco-2 monolayers and found that up to 5 mg/ml concentration, these nanotubes are not cytotoxic and not able to permeate through the intestinal cell layer. Transmission electron microscopic experiments showed that titanate nanotubes are not taken up by cells, only caused a high-density granulation on the surface of the endoplasmic reticulum. According to these results, titanate nanotubes are suitable systems for intestinal drug delivery.

Synergistic effects of cisplatin chemotherapy and gold nanorod-mediated hyperthermia on ovarian cancer cells and tumors.

AIM: The synergistic effects of gold nanorod (GNR)-mediated mild hyperthermia (MHT; 42-43°C) and cisplatin (CP) activity was evaluated against chemoresistant SKOV3 cells in vitro and with a tumor xenograft model. MATERIALS & METHODS: In vitro studies were performed using CP at cytostatic concentrations (5 µM) and polyethylene glycol-stabilized GNRs, using near-infrared laser excitation for MHT. RESULTS: The amount of polyethylene glycol-GNRs used for environmental MHT was 1 µg/ml, several times lower than the loadings used in tumor tissue ablation. GNR-mediated MHT increased CP-mediated cytotoxicity by 80%, relative to the projected additive effect, and flow cytometry analysis suggested MHT also enhanced CP-induced apoptosis. In a pilot in vivo study, systemically administered polyethylene glycol-GNRs generated sufficient levels of MHT to enhance CP-induced reductions in tumor volume, despite their heterogeneous distribution in tumor tissue. CONCLUSION: These studies imply that effective chemotherapies can be developed in combination with low loadings of nanoparticles for localized MHT. Original submitted 6 July 2013; Revised submitted 20 October 2013.

Clay nanotube encapsulation for functional biocomposites.

Natural halloysite clay nanotubes with 50 nm outer- and 15 nm inner- diameters are described as miniature vehicles for sustained release of drugs and proteins. The release time may be adjusted from 10 to 200 h with the tube surface polymeric coating. An explanation of sustained release through locking electrical potential at the nanotube ends is suggested. These biocompatible ceramic tubes may be also used for architectural construction of nanoshells on microbes through alternation with polycations to enhance the intrinsic properties of biological cells. Halloysite nanotubes (pristine or drug-loaded) are well mixable with polar and low-polar polymers allowing for functional biocomposites with enhanced mechanical strength, adhesivity and slow release of drugs or other chemical agents. Halloysite is nontoxic abundantly available from natural deposit material which does not require exfoliation or other complicated energy consuming processing.

Dispersion method for safety research on manufactured nanomaterials.

Nanomaterials tend to agglomerate in aqueous media, resulting in inaccurate safety assessment of the biological response to these substances. The present study searched for suitable dispersion methods for the preparation of nanomaterial suspensions. Titanium dioxide (TiO2) and zinc oxide (ZnO) nanoparticles were dispersed in a biocompatible dispersion medium by direct probe-type sonicator and indirect cup-type sonicator. Size characterization was completed using dynamic light scattering and transmission electron microscopy. A series of dispersion time and output power, as well as two different particle concentrations were tested. Microscopic contamination of metal titanium that broke away from the tip of the probe into the suspension was found. Size of agglomerated nanoparticles decreased with increase in sonication time or output power. Particle concentration did not show obvious effect on size distribution of TiO2 nanoparticles, while significant reduction of secondary diameter of ZnO was observed at higher concentration. A practicable protocol was then adopted and sizes of well-dispersed nanoparticles increased by less than 10% at 7 d after sonication. Multi-walled carbon nanotubes were also well dispersed by the same protocol. The cup-type sonicator might be a useful alternative to the traditional bath-type sonicator or probe-type sonicator based on its effective energy delivery and assurance of suspension purity.

[Environmental air pollutants and the risk of cancer].

The increased combustion of fossil fuels is one of the main reasons for the hazardous changes in the atmospheric composition. The sources of air pollution in urban areas include diesel motor vehicles, residential wood burning, and certain industrial processes. The types of air pollution include gases(eg, carbon monoxide, sulfur dioxide, nitrogen oxides, ozone)and suspended particulate matter(PM)such as PM2.5 and PM10 in diesel exhaust particles. PM2.5 refers to particles less than 2.5 micrometers in diameter. Long-term exposure to PM2.5 can increase the cardiovascular disease risk and lung cancer mortality. Although the role of PM2.5 in the etiology of lung cancer is not very clear, some researchers have shown evidence of increases in lung cancer mortality associated with exposure to PM2.5. Asbestos is also an important cause of cancer of the respiratory tract, particularly lung cancer and mesothelioma. The oncogenic hazards of asbestos fiber have been noted in cases of lowdose environmental exposure, as well in cases of high-dose occupational exposure. The use of asbestos has been strictly prohibited in Japan since 2006. However, large-scale natural disasters such as earthquakes, tsunamis, and typhoons can destroy many buildings and houses that were constructed before the ban on asbestos was initiated, thus resulting in the exposure of human beings to asbestos fibers. In the Cappadocian villages of Tuzkoy, Karain, and Sarihidir in Turkey, 50% of all deaths among villagers are caused by mesothelioma. This condition has been attributed to exposure to erionite, which is a type of fibrous zeolite mineral commonly found in this area of Turkey. However, pedigree studies of these villages showed that mesothelioma was prevalent in certain families but not in others, and that erionite exposure typically causes mesothelioma in those with a genetic predisposition to this disease. Recently, the germline BAP1 mutation was demonstrated in 2 different familial clusters of mesothelioma in the US.

Trafficking of gold nanorods in breast cancer cells: uptake, lysosome maturation, and elimination.

Gold nanorods (AuNRs) have been largely investigated driven by their promising potentials in drug delivery, imaging, and photodynamic therapy because of their distinctive physicochemical properties. It is widely known that AuNRs can be taken up by different cells, however, the trafficking of the nanorods in cells are less known. In this work, the behaviors and fate of AuNRs in the human breast cancer cell line MDA-MB-231 were intensively probed by transmission electron microscopy (TEM) with detailed time resolution, together with induced couple plasmon mass spectroscopy (ICP-MS), confocal microscopy, Western blot, and cell viability assay. We reveal that AuNRs enter the classic lysosome maturation through endocytosis and are sequestered in the vesicular system even during cell division. AuNRs can escape from the lysosomes occasionally and the escaped AuNRs are recycled back into the lysosomal system through cytoprotective autophagy. The dilution of AuNRs in cells is mainly attributed to the cell division rather than exocytosis, because expelled AuNRs can be re-endocytosed by the cells. The feature of vesicular restriction guarantees other organelles such as mitochondria and nucleus are exempted from the direct exposure to AuNRs.

A galvanic replacement route to prepare strongly fluorescent and highly stable gold nanodots for cellular imaging.

Fluorescent gold nanodots (GNDs) are an important kind of nanoprobes. Herein, the application of galvanic replacement for the preparation of fluorescent GNDs is reported. Using presynthesized and size-controlled Ag nanodots (Ag NDs) as templates, the as-prepared GNDs have strong fluorescence (quantum yields ~10%) with high stability and surface bioactivity. The resultant GNDs show excellent photoluminescence properties with high photo-, time-, metal-, and pH-stability, which are attributed to the protective surface layer of glutathione (GSH) and the presence of Au(I)-S complexes on the surface of the gold core. GSH, a naturally occurring and readily available tripeptide with carboxyl and amino functional groups, allows good dispersion of the as-prepared GNDs in aqueous solution and favorable biocompatibility. These advantages, combined with their small size, mean that the as-prepared GNDs have potential application in biological labeling, especially as a DNA probe for the specific detection of nucleic acids. In this study, the CAL-27 cells are used as a model to evaluate the fluorescence imaging of GNDs.

Enhanced single strand breaks of supercoiled DNA in a matrix of gold nanotubes under X-ray irradiation.

Single-strand-breaks (SSBs) of supercoiled DNA (scDNA) molecules were used to probe the enhancement of X-ray radiation effect on scDNA mixed with gold nanotubes (AuNTs) in water. The amounts of measured enhancements using SSBs were significantly lower than the expected increase in energy deposition in water by AuNTs under hard X-ray irradiation. Three factors were identified to negatively affect the enhancement: (1) Attenuation of kinetic energies carried by electrons escaped from AuNTs, (2) Scavenging of OH radicals (˙OH) by the surface of bare AuNTs, and (3) Steric effect due to soluble scDNA molecules away from the surface of AuNTs. Benefits and limits of using gold nanomaterials as radiation enhancers and contrast agents are discussed.

Effects of nanoparticle surface-coupled peptides, functional endgroups, and charge on intracellular distribution and functionality of human primary reticuloendothelial cells.

The medical use of nanoparticles (NPs) has to consider their interactions with the cells of the reticuloendothelial system. In this study the authors used gold nanorods coated by PEG chains bearing peptides or charged functional groups to study their influence on the uptake, subcellular distribution, and activation of human primary reticuloendothelial cells: monocytes, macrophages (MΦ), immature and mature dendritic cells (DC), and endothelial cells (EC). We found that beside MΦ and immature DC also EC internalize large quantities of NPs and observed an increased uptake of positively charged particles. Most notably, NPs accumulated in the MHC II compartment in mature DC that is involved in antigen processing. Furthermore, surface-coupled peptide sequences RGD and GLF altered the activation profile of DC, and modulated cytokine release in both DC and MΦ in a cell specific manner. These data suggest that the charge of NPs mainly influences their uptake, whereas conjugated peptides alter cell functions. FROM THE CLINICAL EDITOR: In this paper the interactions between RES cells and nanoparticles is investigated, concluding that in the case of gold nanorods charge determines uptake characteristics, whereas conjugated peptides determine their function.

Photothermal therapy in a murine colon cancer model using near-infrared absorbing gold nanorods.

The photothermal ablation of solid tumors using exogenous, near-infrared (NIR)-absorbing nanoparticles has been previously investigated using various preclinical models and is currently being evaluated in the clinic. Here, we evaluate the circulation kinetics, preliminary toxicity, and efficacy of photothermal ablation of solid tumors using gold nanorods systemically delivered and passively accumulated in a murine subcutaneous colon cancer model. Tumored animals were infused with nanorods followed by the percutaneous illumination of the tumor with an 808-nm laser. Control groups consisted of laser-only, nanorod-only, and untreated tumored animals. The survival of the treated and control groups were monitored for 60 days post-treatment. The survival of the photothermally treated group was statistically longer than the control groups, with approximately 44% tumor free through the evaluation period. Histopathology of the major organs of animals infused with nanorods did not indicate any significant toxicity at 60 days post-treatment. Particle biodistribution was evaluated by elemental analysis of the major organs of untumored mice at 1, 7, and 30 days after infusion with nanorods. Elemental analysis indicates nanorod clearance from the blood and retention by the reticuloendothelial system. This study indicates that gold nanorods are promising agents for photothermal ablation of solid tumors.